Campos Fernando, Carriel Víctor, Bermejo-Casares Fabiola, Chato-Astrain Jesús, Alaminos Miguel
Tissue Engineering Group, Department of Histology, University of Granada, Granada, Spain.
Instituto de Investigacion Biosanitaria ibs.GRANADA, Granada, Spain.
Methods Mol Biol. 2025;2922:249-265. doi: 10.1007/978-1-0716-4510-9_19.
A thorough evaluation and characterization of advanced therapy medicinal products (ATMPs), including tissue-engineered skin substitutes (TESS), is an essential requirement of the European and National Medicines Agencies. Although clear regulatory criteria have been established for different parameters such as stability, sterility or viability, the histological characterization of bioengineered tissues remains inadequately defined. Histological analyses are crucial for assessing both the in vitro features and in vivo behavior of TESS. This chapter describes the essential histological methods used for assessing TESS, including hematoxylin and eosin staining, and several methods focused on the analysis of specific components of the skin cells and extracellular matrix, such as Picrosirius Red for collagen fibers, Fontana Masson Picrosirius for melanin and collagen, Verhoeff for elastic fibers, Alcian Blue for proteoglycans, and Periodic acid-Schiff for glycoproteins. Relevant immunohistochemical methods for the identification of some specific markers are also included in this chapter. These methods provide a comprehensive practical approach to assessing both in vitro and in vivo histological features and evolution of TESS.
对包括组织工程皮肤替代物（TESS）在内的先进治疗药物产品（ATMPs）进行全面评估和特性描述，是欧洲和各国药品监管机构的一项基本要求。尽管已针对稳定性、无菌性或活力等不同参数制定了明确的监管标准，但生物工程组织的组织学特性描述仍不够明确。组织学分析对于评估TESS的体外特征和体内行为至关重要。本章介绍了用于评估TESS的基本组织学方法，包括苏木精和伊红染色，以及几种专注于分析皮肤细胞和细胞外基质特定成分的方法，如用于胶原纤维的天狼星红染色、用于黑色素和胶原的福尔塔纳·马森天狼星红染色、用于弹性纤维的韦尔霍夫染色、用于蛋白聚糖的阿尔辛蓝染色以及用于糖蛋白的过碘酸希夫染色。本章还包括用于鉴定某些特定标志物的相关免疫组织化学方法。这些方法为评估TESS的体外和体内组织学特征及演变提供了一种全面的实用方法。
